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dUTP used for PCR - (Aug/12/2001 )

I had used the dTTP to do the PCR before, but now I had changed to use the dUTP and the PCR product can not be generated. Does any one know how the PCR can work which the nucleotide used is dUTP.

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You have to be sure that the thermostable polymerase that you're using will incorporate dUTP into DNA, some enzymes like Pfx polymerase will not. I would also suggest using double the concentrations of dUTP in your reactions since Taq tends to have a lower affinity for dUTP. Good luck.

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I have the same problem ... I can't get PCR products for AT rich sequences using dUTP instead of dTTP
I tried
* lowering annealing temperature
* Increasing dUTP concentration (twice as high as normal dNTPs)
* Trying different enzymes

Does anybody knows about an enzyme (or something else) that may solve this PCR problem?

Kind regards
Tim

-stakie-